Systematic conservation planning is well suited to address the many large-scale biodiversity conservation challenges facing the Appalachian region. However, broad, well-connected landscapes will be required to sustain many of the natural resources important to this area into the future. If these landscapes are to be resilient to impending change, it will likely require an orchestrated and collaborative effort reaching across jurisdictional and political boundaries. The first step in realizing this vision is prioritizing discrete places and actions that hold the greatest promise for the protection of biodiversity. The irreplacebility of the landscape was assessed to determine the importance of conservation. The number of target species within a certain area throughout the Appalachian landscape were identified to assist with the identification of important biological areas in the region.